Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 31 AUG 2004 to 7 FEB 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: guideline study (OECD TG 407)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Guidelines for Screening, Toxicity Testing of Chemicals: testing Methods for new Substances (1986)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- HanBrl:WIST (SPF)
- Source: RCC Ltd, Laboratory Animal Services, Füllinsdorf, Switzerland
- Age at study initiation: 7 weeks
- Weight range (and mean) at acclimatization: Males 141-154 g (mean 147.9); Females 119- 134 g (mean: 126.1 g)
- Housing: in groups of 5 in macrolon cages (type 4)
- Diet: Provimi Kliba 3433 rat maintenance diet (batch nos. 34/04, 42/04, 53/04; Provimi Kliba AG, Kaiseraugst, Switzerland), ad libitum
- Water: community tap-water from Itingen (Switzerland), ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30 to 70
- Air changes (per hr): 10 to 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- dose formulations were prepared daily at room temperature and homogeneity was kept using a magnetic stirrer.
- the test item is stable in dose formulations for up to 7 days

VEHICLE
- polyethylene glycol 300
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration, homogeneity and stability (after 2 hours and 7 days) of the dose formulations were determined in samples taken after experimental start. Concentration and homogeneity of the dose formulations were determined in samples taken during week 3 of the treatment.
Analysis was performed spectrophotometrically by UUV/VIS-detection at 530 nm by RCC Ltd according to amethod provided by the sponsor.
Duration of treatment / exposure:
28 days
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 50, 200, 1000 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
- 5 male and 5 female in each dose group
- 5 male and 5 female for recovery groups at dose 0 and 1000 mg/ kg bw/day
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on the results of a dose range finding study with the test item (5 days oral exposure of male and female rats to 200, 600 and 1000 mg/kg bw/day did not cause changes in clinical appearance, body weights, food consumption, organ weights and macroscopic examination (only in one female control animal there was pelvic dilatation in the kidney on the left side) and all animals survived until scheduled necropsy)
- Post-exposure recovery period in satellite groups: the duration of recovery was 14 days
Positive control:
- none

Examinations

Observations and examinations performed and frequency:
MORTALITY/VIABILITY: Yes
- twice daily

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once before treatment, twice daily on days 1-3, once daily on days 4-28 and once daily during days 29-42 (recovery)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before treatment and thereafter once weekly

BODY WEIGHT: Yes, weekly

FOOD CONSUMPTION: yes, weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

HAEMATOLOGY: Yes
- Parameters checked see below in Hematology

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks and after 6 weeks (the latter only applied for recovery groups)
- Animals fasted: Yes (appr. 18 hours before blood collection)
- How many animals: all animals
- Parameters checked see below

URINALYSIS: Yes
- Time schedule for collection of urine: after 4 weeks and after 6 weeks (the latter only applied for recovery groups)
- Metabolism cages used for collection of urine: No (urine was collected during the 18-hour fasting period into a specimen vial)
- Animals fasted: Yes
- Parameters checked see below

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: during week 4
- Dose groups that were examined: all
- Battery of functions tested: grip strength / locomotor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (parameters checked see below)
HISTOPATHOLOGY: Yes (parameters checked see below)
Statistics:
- Dunnett-test
- Fisher's exact test (applied on macroscopic findings)
- clinical laboratory data were analysed via ANOVA according to Bartlett or non-parametric Kruskal-Wallis test, treated groups were compared to control groups using ANOVA and Dunnett's test or combining Kruskal-Wallis test with the Dunn's test

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY:
- no deaths occurred during the test
- two males treated with 1000 mg/kg/day presented a temporary hair loss during week 2 of treatment with the test item
- one female showed respiratory rales during two days of treatment with the test item
- dark red feces were noted in animals treated with the test item ("grade 1" in animlas treated with 50 or 200 mg/kg/day; "grade 2" in animals treated with 1000 mg/kg/day), which stopped at the termination of the treatment
- overall no clinical observations of toxicological relevance were found

BODY WEIGHT AND WEIGHT GAIN
No test item-related changes of toxicological relevance were noted in the mean body weights and body weight gain, at any dose level, when compared with controls, after four weeks of treatment and after recovery.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The mean daily food consumption and the relative food consumption of the test item-treated males and females compared favorably with their respective controls during the treatment and the recovery periods.

CLINICAL LABORATORY INVESTIGATOIONS
-HAEMATOLOGY
No changes of toxicological relevance were observed in the hematology parameters after the treatment or recovery periods in both males and females.

-CLINICAL CHEMISTRY
No changes of toxicological relevance were noted in the clinical biochiemistry parameters after the treatment and the recovery periods.

-URINALYSIS
No changes of toxicological relevance were observed in the urinalysis parameters after the treatment with the test item and the recovery period in both males and females, at any dose level tested.

ORGAN WEIGHTS
No test item related differences in the mean absolute or relative organ weights were noted after four weeks' treatment or after two weeks' recovery in males or female rats of any dose level.

GROSS PATHOLOGY
- The pink coloured and birefringent granules were recorded in the lumen of the gastrointestinal tract of most animals of the high dose group (1000 mg/kg/d). Without any tissue reaction these particles were regarded to present residues of the adminsitered test item. At the end of the recovery period the material was no longer present.

HISTORICAL CONTROL DATA (if applicable)
- all microscopic findings recorded did not distinguish significantly treated rats from control rats and were considered to be spontaneous in nature and within the normal background pathology commonly seen in rats of this strain and age.

OTHER FINDINGS
-transitory increase in the mean locomotor activity was observed in males and females treated with 1000 mg/kg/d during one measurement interval. In the absence of a clear dose-response relationship, these changes were considered as incidental.

Effect levels

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: no clinical signs; no mortality; body weight development was not impaired; food consumption was not affected; no findings in haematology; no effects found in clinical chemistry, urinalysis, gross pathology and histopathology

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
There were no changes in clinical appearance, functional observations, body weights, food consumption, clinical laboratory investigations, macroscopic examination, organ weights and microscopic examination that were considered to be an effect of treatment.
From the results presented in this report a definitive No Observed Adverse Effect Level (NOAEL) for PIGMENT RED 282 of 1000 mg/kg/day was established.
Executive summary:

A 28 -day repeated dose toxicity study with PIGMENT RED 282 administered by oral gavage to Wistar rats (SPF-bred; 5/sex/dose) was performed according to OECD TG 407. The dose levels for this study were 50, 200 and 1000 mg/kg bw/day. A control group was treated similarly with the vehicle, PEG 300, only. The dosing lasted for 28 days and a 14 day recovery group for controls and the high dosage group (1000 mg/kg bw/d) was included. The test item revealed no treatment-related findings. From the results presented in this report a definitive No Observed Adverse Effect Level(NOAEL) for the test item of 1000 mg/kg bw/day was established.